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Article ; Online: Whole-Mount Imaging of Adipose Tissue Macrophages.

Sorokin, Lydia / Corrêa, Luis Henrique

Methods in molecular biology (Clifton, N.J.)

2023 Volume 2713, Page(s) 307–322

Abstract: The adipose tissue comprises highly heterogeneous macrophage populations, which play critical roles in the regulation of adipose tissue function and dysfunction during health and disease. Whole-amount staining is a powerful technique for macrophage ... ...

Abstract	The adipose tissue comprises highly heterogeneous macrophage populations, which play critical roles in the regulation of adipose tissue function and dysfunction during health and disease. Whole-amount staining is a powerful technique for macrophage characterization within the 3D environment of the adipose tissue, enabling the visualization of different macrophage populations and their interaction with other cells within their in vivo niche. Due to the high-fat content and softness, freezing and sectioning of adipose tissue is difficult, and distortion of tissue morphology typically occurs, especially in the case of white adipose tissue. We describe here a whole-mount staining alternative for adipose tissue imaging that preserves all structures and allows high-resolution image acquisition. We address in a step-by-step manner how to perform immunofluorescence staining of different fat pads and how to optimally visualize cellular and acellular (extracellular matrix) constituents of the adipose tissue and its vasculature, as well as resident and infiltrating macrophage populations.
MeSH term(s)	Adipose Tissue ; Adipose Tissue, White ; Diagnostic Imaging ; Extracellular Matrix ; Macrophages
Language	English
Publishing date	2023-08-25
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3437-0_21
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Article ; Online: ECM-integrin signalling instructs cellular position sensing to pattern the early mouse embryo.

Kim, Esther Jeong Yoon / Sorokin, Lydia / Hiiragi, Takashi

Development (Cambridge, England)

2022 Volume 149, Issue 1

Abstract: Development entails patterned emergence of diverse cell types within the embryo. In mammals, cells positioned inside the embryo give rise to the inner cell mass (ICM), which eventually forms the embryo itself. Yet, the molecular basis of how these cells ... ...

Abstract	Development entails patterned emergence of diverse cell types within the embryo. In mammals, cells positioned inside the embryo give rise to the inner cell mass (ICM), which eventually forms the embryo itself. Yet, the molecular basis of how these cells recognise their 'inside' position to instruct their fate is unknown. Here, we show that provision of extracellular matrix (ECM) to isolated embryonic cells induces ICM specification and alters the subsequent spatial arrangement between epiblast (EPI) and primitive endoderm (PrE) cells that emerge within the ICM. Notably, this effect is dependent on integrin β1 activity and involves apical-to-basal conversion of cell polarity. We demonstrate that ECM-integrin activity is sufficient for 'inside' positional signalling and is required for correct EPI/PrE patterning. Thus, our findings highlight the significance of ECM-integrin adhesion in enabling position sensing by cells to achieve tissue patterning.
MeSH term(s)	Animals ; Body Patterning ; Cell Differentiation ; Cell Polarity ; Cells, Cultured ; Ectoderm/cytology ; Ectoderm/metabolism ; Endoderm/cytology ; Endoderm/metabolism ; Extracellular Matrix/metabolism ; Integrin beta1/metabolism ; Mice ; Mice, Inbred C57BL ; Mouse Embryonic Stem Cells/cytology ; Mouse Embryonic Stem Cells/metabolism ; Signal Transduction
Chemical Substances	Integrin beta1
Language	English
Publishing date	2022-01-13
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	90607-4
ISSN	1477-9129 ; 0950-1991
ISSN (online)	1477-9129
ISSN	0950-1991
DOI	10.1242/dev.200140
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Article ; Online: The impact of the extracellular matrix on inflammation.

Sorokin, Lydia

Nature reviews. Immunology

2010 Volume 10, Issue 10, Page(s) 712–723

Abstract: The advent of in situ immunology and intravital analyses of leukocyte movement in tissues has drawn attention to the previously neglected extracellular matrix (ECM) and its role in modulating immune cell behaviour in inflamed tissues. The ECM exists in ... ...

Abstract	The advent of in situ immunology and intravital analyses of leukocyte movement in tissues has drawn attention to the previously neglected extracellular matrix (ECM) and its role in modulating immune cell behaviour in inflamed tissues. The ECM exists in different biochemical and structural forms; both their individual components and three-dimensional ultrastructure impart specific signals to cells that modulate basic functions that are important for the early steps in inflammation, such as immune cell migration into inflamed tissues and immune cell differentiation. In chronically inflamed tissues, aberrant ECM expression and fragments of the ECM that are derived from tissue-remodelling processes can influence immune cell activation and survival, thereby actively contributing to immune responses at these sites.
MeSH term(s)	Animals ; Extracellular Matrix/immunology ; Humans ; Inflammation/immunology
Language	English
Publishing date	2010-09-24
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2062776-2
ISSN	1474-1741 ; 1474-1733
ISSN (online)	1474-1741
ISSN	1474-1733
DOI	10.1038/nri2852
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Article ; Online: The extracellular matrix of lymph node reticular fibers modulates follicle border interactions and germinal center formation.

Song, Jian / Deshpande, Tushar / Zhang, Xueli / Hannocks, Melanie-Jane / Lycke, Nils / Cardell, Susanna L / Sorokin, Lydia

iScience

2023 Volume 26, Issue 5, Page(s) 106753

Abstract: Germinal center (GC) formation and antibody production in lymph node follicles require coordinated interactions between B-cells, T-cells and dendritic cells (DCs), orchestrated by the extracellular matrix-rich reticular fiber (RF) network. We describe a ... ...

Abstract	Germinal center (GC) formation and antibody production in lymph node follicles require coordinated interactions between B-cells, T-cells and dendritic cells (DCs), orchestrated by the extracellular matrix-rich reticular fiber (RF) network. We describe a unique laminin 523-containing RF network around and between follicles that associates with PDGFrecβ
Language	English
Publishing date	2023-04-28
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2023.106753
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Article ; Online: The niche matters: origin, function and fate of CNS-associated macrophages during health and disease.

Dalmau Gasull, Adrià / Glavan, Martina / Samawar, Sai K Reddy / Kapupara, Kishan / Kelk, Joe / Rubio, Marina / Fumagalli, Stefano / Sorokin, Lydia / Vivien, Denis / Prinz, Marco

Acta neuropathologica

2024 Volume 147, Issue 1, Page(s) 37

Abstract: There are several cellular and acellular structural barriers associated with the brain interfaces, which include the dura, the leptomeninges, the perivascular space and the choroid plexus epithelium. Each structure is enriched by distinct myeloid ... ...

Abstract	There are several cellular and acellular structural barriers associated with the brain interfaces, which include the dura, the leptomeninges, the perivascular space and the choroid plexus epithelium. Each structure is enriched by distinct myeloid populations, which mainly originate from erythromyeloid precursors (EMP) in the embryonic yolk sac and seed the CNS during embryogenesis. However, depending on the precise microanatomical environment, resident myeloid cells differ in their marker profile, turnover and the extent to which they can be replenished by blood-derived cells. While some EMP-derived cells seed the parenchyma to become microglia, others engraft the meninges and become CNS-associated macrophages (CAMs), also referred to as border-associated macrophages (BAMs), e.g., leptomeningeal macrophages (MnMΦ). Recent data revealed that MnMΦ migrate into perivascular spaces postnatally where they differentiate into perivascular macrophages (PvMΦ). Under homeostatic conditions in pathogen-free mice, there is virtually no contribution of bone marrow-derived cells to MnMΦ and PvMΦ, but rather to macrophages of the choroid plexus and dura. In neuropathological conditions in which the blood-brain barrier is compromised, however, an influx of bone marrow-derived cells into the CNS can occur, potentially contributing to the pool of CNS myeloid cells. Simultaneously, resident CAMs may also proliferate and undergo transcriptional and proteomic changes, thereby, contributing to the disease outcome. Thus, both resident and infiltrating myeloid cells together act within their microenvironmental niche, but both populations play crucial roles in the overall disease course. Here, we summarize the current understanding of the sources and fates of resident CAMs in health and disease, and the role of the microenvironment in influencing their maintenance and function.
MeSH term(s)	Mice ; Animals ; Proteomics ; Macrophages/pathology ; Central Nervous System/pathology ; Microglia ; Meninges
Language	English
Publishing date	2024-02-12
Publishing country	Germany
Document type	Journal Article ; Review
ZDB-ID	1079-0
ISSN	1432-0533 ; 0001-6322
ISSN (online)	1432-0533
ISSN	0001-6322
DOI	10.1007/s00401-023-02676-9
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Article ; Online: Endothelial cells and macrophages as allies in the healthy and diseased brain.

Denes, Adam / Hansen, Cathrin E / Oezorhan, Uemit / Figuerola, Sara / de Vries, Helga E / Sorokin, Lydia / Planas, Anna M / Engelhardt, Britta / Schwaninger, Markus

Acta neuropathologica

2024 Volume 147, Issue 1, Page(s) 38

Abstract: Diseases of the central nervous system (CNS) are often associated with vascular disturbances or inflammation and frequently both. Consequently, endothelial cells and macrophages are key cellular players that mediate pathology in many CNS diseases. ... ...

Abstract	Diseases of the central nervous system (CNS) are often associated with vascular disturbances or inflammation and frequently both. Consequently, endothelial cells and macrophages are key cellular players that mediate pathology in many CNS diseases. Macrophages in the brain consist of the CNS-associated macrophages (CAMs) [also referred to as border-associated macrophages (BAMs)] and microglia, both of which are close neighbours or even form direct contacts with endothelial cells in microvessels. Recent progress has revealed that different macrophage populations in the CNS and a subset of brain endothelial cells are derived from the same erythromyeloid progenitor cells. Macrophages and endothelial cells share several common features in their life cycle-from invasion into the CNS early during embryonic development and proliferation in the CNS, to their demise. In adults, microglia and CAMs have been implicated in regulating the patency and diameter of vessels, blood flow, the tightness of the blood-brain barrier, the removal of vascular calcification, and the life-time of brain endothelial cells. Conversely, CNS endothelial cells may affect the polarization and activation state of myeloid populations. The molecular mechanisms governing the pas de deux of brain macrophages and endothelial cells are beginning to be deciphered and will be reviewed here.
MeSH term(s)	Endothelial Cells ; Brain/pathology ; Macrophages ; Central Nervous System/pathology ; Microglia
Language	English
Publishing date	2024-02-12
Publishing country	Germany
Document type	Journal Article ; Review
ZDB-ID	1079-0
ISSN	1432-0533 ; 0001-6322
ISSN (online)	1432-0533
ISSN	0001-6322
DOI	10.1007/s00401-024-02695-0
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Article ; Online: Fractone Bulbs Derive from Ependymal Cells and Their Laminin Composition Influence the Stem Cell Niche in the Subventricular Zone.

Nascimento, Marcos Assis / Sorokin, Lydia / Coelho-Sampaio, Tatiana

The Journal of neuroscience : the official journal of the Society for Neuroscience

2018 Volume 38, Issue 16, Page(s) 3880–3889

Abstract: Fractones are extracellular matrix structures in the neural stem cell niche of the subventricular zone (SVZ), where they appear as round deposits named bulbs or thin branching lines called stems. Their cellular origin and what determines their ... ...

Abstract	Fractones are extracellular matrix structures in the neural stem cell niche of the subventricular zone (SVZ), where they appear as round deposits named bulbs or thin branching lines called stems. Their cellular origin and what determines their localization at this site is poorly studied, and it remains unclear whether they influence neural stem and progenitor cell formation, proliferation, and/or maintenance. To address these questions, we analyzed whole-mount preparations of the lateral ventricle of male and female mice by confocal microscopy using different extracellular matrix and cell markers. We found that bulbs are rarely connected to stems and that they contain laminin α5 and α2 chains, respectively. Fractone bulbs were profusely distributed throughout the SVZ and appeared associated with the center of pinwheels, a critical site for adult neurogenesis. We demonstrate that bulbs appear at the apical membrane of ependymal cells at the end of the first week after birth. The use of transgenic mice lacking laminin α5 gene expression (
MeSH term(s)	Animals ; Cell Proliferation ; Ependyma/cytology ; Ependyma/metabolism ; Extracellular Matrix/metabolism ; Female ; Forkhead Transcription Factors/genetics ; Forkhead Transcription Factors/metabolism ; Laminin/genetics ; Laminin/metabolism ; Lateral Ventricles/cytology ; Lateral Ventricles/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Neural Stem Cells/cytology ; Neural Stem Cells/metabolism ; Stem Cell Niche
Chemical Substances	FOXJ1 protein, mouse ; Forkhead Transcription Factors ; Laminin
Language	English
Publishing date	2018-03-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	604637-x
ISSN	1529-2401 ; 0270-6474
ISSN (online)	1529-2401
ISSN	0270-6474
DOI	10.1523/JNEUROSCI.3064-17.2018
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Article: Superoxide Dismutase-3 Downregulates Laminin α5 Expression in Tumor Endothelial Cells via the Inhibition of Nuclear Factor Kappa B Signaling.

Carmona-Rodríguez, Lorena / Martínez-Rey, Diego / Martín-González, Paula / Franch, Mónica / Sorokin, Lydia / Mira, Emilia / Mañes, Santos

Cancers

2022 Volume 14, Issue 5

Abstract: The balance between laminin isoforms containing the α5 or the α4 chain in the endothelial basement membrane determines the site of leukocyte diapedesis under inflammatory conditions. Extracellular superoxide dismutase (SOD3) induces laminin α4 expression ...

Abstract	The balance between laminin isoforms containing the α5 or the α4 chain in the endothelial basement membrane determines the site of leukocyte diapedesis under inflammatory conditions. Extracellular superoxide dismutase (SOD3) induces laminin α4 expression in tumor blood vessels, which is associated with enhanced intratumor T cell infiltration in primary human cancers. We show now that SOD3 overexpression in neoplastic and endothelial cells (ECs) reduces laminin α5 in tumor blood vessels. SOD3 represses the laminin α5 gene (
Language	English
Publishing date	2022-02-26
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers14051226
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Article ; Online: Single-cell transcriptomics reveals functionally specialized vascular endothelium in brain.

Jeong, Hyun-Woo / Diéguez-Hurtado, Rodrigo / Arf, Hendrik / Song, Jian / Park, Hongryeol / Kruse, Kai / Sorokin, Lydia / Adams, Ralf H

eLife

2022 Volume 11

Abstract: The blood-brain barrier (BBB) limits the entry of leukocytes and potentially harmful substances from the circulation into the central nervous system (CNS). While BBB defects are a hallmark of many neurological disorders, the cellular heterogeneity at the ...

Abstract	The blood-brain barrier (BBB) limits the entry of leukocytes and potentially harmful substances from the circulation into the central nervous system (CNS). While BBB defects are a hallmark of many neurological disorders, the cellular heterogeneity at the neurovascular interface, and the mechanisms governing neuroinflammation are not fully understood. Through single-cell RNA sequencing of non-neuronal cell populations of the murine cerebral cortex during development, adulthood, ageing, and neuroinflammation, we identify reactive endothelial venules, a compartment of specialized postcapillary endothelial cells that are characterized by consistent expression of cell adhesion molecules, preferential leukocyte transmigration, association with perivascular macrophage populations, and endothelial activation initiating CNS immune responses. Our results provide novel insights into the heterogeneity of the cerebral vasculature and a useful resource for the molecular alterations associated with neuroinflammation and ageing.
MeSH term(s)	Animals ; Blood-Brain Barrier/metabolism ; Brain/metabolism ; Cell Adhesion Molecules/metabolism ; Endothelial Cells/metabolism ; Endothelium, Vascular/metabolism ; Mice ; Transcriptome
Chemical Substances	Cell Adhesion Molecules
Language	English
Publishing date	2022-10-05
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.57520
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Article ; Online: Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function.

Burmeister, Miriam / Fraunenstein, Annika / Kahms, Martin / Arends, Laura / Gerwien, Hanna / Deshpande, Tushar / Kuhlmann, Tanja / Gross, Catharina C / Naik, Venu N / Wiendl, Heinz / Klingauf, Juergen / Meissner, Felix / Sorokin, Lydia

Science advances

2023 Volume 9, Issue 29, Page(s) eadg0686

Abstract: The gelatinases, matrix metalloproteinase 2 (MMP-2) and MMP-9, are key for leukocyte penetration of the brain parenchymal border in neuroinflammation and the functional integrity of this barrier; however, it is unclear which MMP substrates are involved. ... ...

Abstract	The gelatinases, matrix metalloproteinase 2 (MMP-2) and MMP-9, are key for leukocyte penetration of the brain parenchymal border in neuroinflammation and the functional integrity of this barrier; however, it is unclear which MMP substrates are involved. Using a tailored, sensitive, label-free mass spectrometry-based secretome approach, not previously applied to nonimmune cells, we identified 119 MMP-9 and 21 MMP-2 potential substrates at the cell surface of primary astrocytes, including known substrates (β-dystroglycan) and a broad spectrum of previously unknown MMP-dependent events involved in cell-cell and cell-matrix interactions. Using neuroinflammation as a model of assessing compromised astroglial barrier function, a selection of the potential MMP substrates were confirmed in vivo and verified in human samples, including vascular cell adhesion molecule-1 and neuronal cell adhesion molecule. We provide a unique resource of potential MMP-2/MMP-9 substrates specific for the astroglia barrier. Our data support a role for the gelatinases in the formation and maintenance of this barrier but also in astrocyte-neuron interactions.
MeSH term(s)	Humans ; Gelatinases/metabolism ; Matrix Metalloproteinase 2/metabolism ; Matrix Metalloproteinase 9/metabolism ; Blood-Brain Barrier/metabolism ; Astrocytes/metabolism ; Neuroinflammatory Diseases
Chemical Substances	Gelatinases (EC 3.4.24.-) ; Matrix Metalloproteinase 2 (EC 3.4.24.24) ; Matrix Metalloproteinase 9 (EC 3.4.24.35)
Language	English
Publishing date	2023-07-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2810933-8
ISSN	2375-2548 ; 2375-2548
ISSN (online)	2375-2548
ISSN	2375-2548
DOI	10.1126/sciadv.adg0686
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